The Marshall University Imaging Core will support the imaging activities of five junior investigators and potential pilot investigators as part of the project titled Appalachian Center for Cellular transport in Obesity Related Disorders (ACCORD). The five junior investigators have identified abnormalities in cellular transport related to disease states which are particularly associated with obesity, including cancer, hypertension and skeletal malformations. With the long-range goal of minimizing the health impact of dysregulated transport, or even blocking or reversing processes related to these transport mechanisms, these researchers seek to develop molecular level descriptions, which will lead to an understanding of the transport mechanism and the mechanism by which external factors can affect transport. The Imaging Core will support the long-range goals of these investigators by providing visualization tools and techniques which will enable them to determine the relative numbers and distributions of the specific proteins they have determined are associated with transport anomalies in selected disease states. The principle imaging tool which will be applied for the visualization tasks is a Multiphoton Fluorescence Microscope, which will enable high resolution optical imaging of tagged proteins. The multiphoton capability increases the quality of optical imaging in tissue, and will be used in studies comparing normal and abnormal tissues. This imaging capability will be enhanced, for researchers developing suitable model systems, via the production of custom tagged proteins suitable for use in live cell imaging. Live cell imaging will provide the opportunity to observe changes in protein concentrations and/or distributions within a cell occurring in response to a physiologically relevant cue or resulting from introduction of a pharmacological agent. Techniques enabled by fluorescent protein tagging, including FRET (fluorescence resonant energy transfer) and FRAP (fluorescence recovery after photobleaching) may be employed to test developing hypotheses of protein transport mechanisms and their anomalies. Additional capabilities, including single molecule fluorescence imaging and atomic force microscopy will be available for addressing evolving project requirements.